Rho激酶抑制剂Y-27632对人牙髓干细胞增殖能力的影响

doi:10.13701/j.cnki.kqyxyj.2016.03.016

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (3): 278-281.DOI: 10.13701/j.cnki.kqyxyj.2016.03.016

Rho激酶抑制剂Y-27632对人牙髓干细胞增殖能力的影响

于天瑶, 潘爽, 何丽娜, 李艳萍, 张琳, 牛玉梅

哈尔滨医科大学口腔医学院牙体牙髓病科黑龙江哈尔滨 150001

收稿日期:2015-08-24 出版日期:2016-03-28 发布日期:2016-03-29
通讯作者: 牛玉梅,E-mail:yumeiniu@163.com
作者简介:于天瑶(1989~ ),女,黑龙江人,医师,硕士在读,主要从事牙体牙髓病的临床治疗工作。
基金资助:
国家自然科学基金(编号:81271132)
黑龙江省教育厅科学技术研究项目(编号:12531234)
黑龙江省自然科学基金项目(编号:HZ01440)

Functional Role of Rho Kinase Inhibitor Y-27632 on Proliferation of Human Dental Pulp Stem Cells.

YU Tian-yao, PAN Shuang, HE Li-na, LI Yan-ping, ZHANG Lin, NIU Yu-mei^*

Department of Endodontics, Stomatology College of Harbin Medical University. Harbin 150001, China

Received:2015-08-24 Online:2016-03-28 Published:2016-03-29

摘要/Abstract

摘要： 目的: 初步研究Rho激酶抑制剂Y-27632对人牙髓干细胞(human dental pulp stem cells,hDPSCs)增殖能力的影响。方法: 采用体外贴壁式培养法培养人牙髓干细胞,应用Rho激酶抑制剂Y-27632,根据培养条件,分为普通培养液培养组(Con)及普通培养液+抑制剂Y-27632培养组(Con+Y),采用MTT法检测两种培养条件下人牙髓干细胞培养24、48、72、96 h细胞活性;采用DAPI染色法及流式细胞定量检测技术(FCM)比较两组在培养48 h的细胞数量及细胞周期分布情况。结果: MTT结果显示,实验组hDPSCs细胞增殖曲线较对照组明显上移,且增殖高峰期提前;培养48 h,DAPI染色结果显示,实验组细胞总数量明显多于对照组;FCM结果显示,对照组G₀/G₁期细胞比例为(66.8±6.84)%,S期细胞比例为(25.17±0.62)%,实验组G0/G1期细胞比例为(58.59±1.76)%,S期细胞比例为(31.34±1.16)%,实验组S期细胞比例明显高于对照组(P<0.05)。 结论: Rho激酶抑制剂Y-27632促进人牙髓干细胞DNA合成、细胞分裂及增殖。

关键词: 人牙髓干细胞, Rho激酶, Y-27632, 增殖

Abstract: Objective: To study the influence of Rho kinase inhibitor Y-27632 on the proliferation of human dental pulp stem cells. Methods: Human dental pulp stem cells (hDPSCs) were seeded in plates，then divided randomly into two groups: cultured with or without Y-27632. Cellular proliferation was determined by DAPI staining and MTT assay. Cell cycle was analyzed by flow cytometry (FCM). Results: After 24, 48, 72 and 96h of cultivation, the optical density (OD) in the Y-27632 treated group was significantly higher than that in the control group. Moreover, the peak time during proliferation was earlier in the treat group. After 48h cultured, compared with the control group, the DAPI staining showed that the proportion of total cells was significantly increased in the Y-27632 treated group. The FCM results indicated that (66.8±6.84)% of the hDPSCs were in G0/G1 phase (25.17±0.62)% in S phase. After Y-27632 treatment, (58.59±1.76)% of the hDPSCs were in G0/G1 phase (31.34±1.16)% in S phase. The cell proliferation during S phase was significantly higher after stimulation by Y-27632 than that in the control group (P<0.05). Conclusion: Inactivation of Rho kinase in the hDPSCs by inhibitor Y-27632 led more hDPSCs to entering into the DNA synthesis stage and promoted the division and proliferation of hDPSCs.

Key words: Human dental pulp stem cells , Rho kinase , Y-27632 , Cell proliferation

中图分类号:

R780.1

于天瑶, 潘爽, 何丽娜, 李艳萍, 张琳, 牛玉梅. Rho激酶抑制剂Y-27632对人牙髓干细胞增殖能力的影响[J]. 口腔医学研究, 2016, 32(3): 278-281.

YU Tian-yao, PAN Shuang, HE Li-na, LI Yan-ping, ZHANG Lin, NIU Yu-mei. Functional Role of Rho Kinase Inhibitor Y-27632 on Proliferation of Human Dental Pulp Stem Cells.[J]. Journal of Oral Science Research, 2016, 32(3): 278-281.

参考文献

[1] Kamoub A E,Symons M,Campbell S L,et al. Molecular basis for rho gtpase signaling specificity [J]. Breast Cancer Res Treat,2004,84(1)∶61-71
[2] Emre N,Vidal JG,Elia J,et al. The Rock inhibitor Y-27632 improves recovery of human embryonic stem cells after fluorescence-activated cell sorting with multiple cell surface markers [J]. Plos S One,2010,5(8)∶e12148
[3] 张巍巍,牛玉梅,等.人牙髓干细胞在聚乳酸-聚乙醇共聚物支架上粘附与增殖的研究[J].口腔医学研究,2011,27(12)∶1083-1088
[4] Nakashina M,Mizunuma K,Murakami T,et al. Induction of dental pulp stem cell differentiation into odontoblast by electroporation-mediated gene delivery of growth/differentiation factor 11(GDF 11) [J]. Gene Ther,2002,9∶814-818
[5] Stokowski A,Shi S,Sun T,et al. EphB/ephrin-B interaction medicates adult stem cell attachment,spreading,and migration:implications for dental tissue repair [J]. Stem cells,2007,5(1)∶156-164
[6] Bement W M,Miller A L,von Dassow G,et al. Rho GTPase activity zones and transient contractile array [J]. Bioessays,2006,28(10)∶983-993
[7] Bhandary L,Whipple RA,Vitolo MI,et al. Rock inhibition promotes microtentacles that enhance reattachment of breast cancer cells [J]. Oncotarget,2015,6(8)∶6251-66
[8] 吴相周,潘爽,牛玉梅,等.模拟微重力对人牙髓干细胞细胞骨架及迁移能力影响的研究[J].口腔医学研究,2015,31(2)∶137-143
[9] Lamas NJ,Serra SC,Salgado AJ,et al. Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells [J]. Stem Cells Cloning,2015,8∶15-26
[10] Otsu K,Kishigami R,Fujiwara N,et al. functional role of rho-kinase in ameloblast differentiation [J]. Cell Physiol,2011,226(10)∶2527-34
[11] 吕刚,姚鑫,等.小分子干扰RNA介导RhoA沉默优化骨髓间充质干细胞培养[J].中国组织工程研究与临床康复,2010,14(45)∶8365-8368
[12] Guo Y,Liu Q,Yang Y,Guo X,et al. The effects of rock inhibitor Y-27632 on injectable spheroids of bovine corneal endothelial cells [J]. Cell Reprogram,2014,10∶77-87

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Rho激酶抑制剂Y-27632对人牙髓干细胞增殖能力的影响

Functional Role of Rho Kinase Inhibitor Y-27632 on Proliferation of Human Dental Pulp Stem Cells.

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